Selection and enrichment of plant protoplast heterokaryons of Brassicaceae by flow sorting

56Citations
Citations of this article
3Readers
Mendeley users who have this article in their library.
Get full text

Abstract

The experimental conditions for an efficient and reproducible enrichment of fusion products by flow cytometry, using protoplasts of different Brassica species as hybridization material, have been investigated. The heterokaryons were identified by the endogenous chlorophyll autofluorescencence of mesophyll protoplasts of one parent and the fluorescense of exogenously supplied carboxyfluorescin to the hypocotyl protoplasts of the other parent. By using a low head drive frequency (11 kHz), a large nozzle (110 μm) and a low nozzle pressure (30-35 kPa) good survival of the protoplasts was obtained after sorting. Heterokaryons were sorted using these parameters and on average 80% of the protoplasts were fusion products as judged by microscopy. They were cultured in small volumes, 150 μl, and started to divide after 3-5 days and regenerated calli easily. Isozyme analysis of the calli confirmed that 81% had the pattern typical for a hybrrid. Differentiation into shoots have been obtained from some of the hybrid calli; these shoots were also confirmed to be true hybrids. © 1986.

Cite

CITATION STYLE

APA

Glimelius, K., Djupsjöbacka, M., & Fellner-Feldegg, H. (1986). Selection and enrichment of plant protoplast heterokaryons of Brassicaceae by flow sorting. Plant Science, 45(2), 133–141. https://doi.org/10.1016/0168-9452(86)90049-X

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free